Автореферат и диссертация по медицине (14.00.25) на тему:Фармакологическая регуляция функции Na#2+#1-K#2+#1-2Cl#2-#1-котранспортера скелетных мышц и изучение его роли в гомеостазе калия и воды

ВВЕДЕНИЕ.

1. ОБЗОР ЛИТЕРАТУРЫ. Роль Ка+-К+-2С1- котранспортера в поддержании гомеостаза калия и воды в клетках млекопитающих

1.1. Роль На+-К+-2СГ-котранспортера в клеточном транспорте калия и воды

1.2. Внутриклеточные механизмы и фармакологическая регуляция активности Ш+-К+-2СГ-котранспортера

1.3. Роль нарушений клеточного транспорта калия и воды скелетных мышц в патогенезе сахарного диабета. Терапевтческое влияние лекарственных препаратов и физической активности

2. МАТЕРИАЛЫ И МЕТОДЫ ИССЛЕДОВАНИЯ.

3. РЕЗУЛЬТАТЫ СОБСТВЕННЫХ ИССЛЕДОВАНИЙ

3.1. Изучение функциональной роли Ка+-К+-2С1-котранспортера в медленной" и "быстрой" скелетных мышцах крысы

3.1.1. Влияние адреноагонистов, инсулина, сократительной активности и гиперосмолярности на буметанид-чувствительный транспорт ЯЬ в скелетной мышце

3.1.2. Влияние изопреналина, инсулина и гиперосмолярности на на буметанид-чувствительный объём мышечных клеток

3.2. Оценка действия адренергических агонистов и инсулина на №+-К+-2СГ-котранспортер-опосредованный транспорт калия в "медленной" и "быстрой" скелетных мышцах крысы

3.2.1. Изучение влияния фармакологических ингибиторов ЕЯК1,2 МАРК на опосредованный изопреналином и фенилэфрином буметанид-чувствительный транспорт калия в скелетной мышце

3.2.2. Изучение влияния фармакологических ингибиторов С^од-иротеинов, ЯаМ, р38 МАРК и фосфатидилинозитол 3-киназы и активаторов ПКА на опосредованный изопреналином буметанид-чувствительный транспорт калия в скелетной мышце

3.2.3. Изучение влияния изопреналина на фосфорилирование ИаМ,

ERK1,2 МАРК и Akt в присутствии фармакологических ингибиторов активности Gi/o/t-протсинов и PI 3-К

3.2.4. Изучение влияния инсулина на буметанид-чувствительный транспорт калия в скелетной мышце. Действие изопреналина и фармакологических ингибиторов п38 МАПК и фосфатидилинозитол 3-киназы

3.2.5. Изучение влияния инсулина на фосфорилирование ERK1,

МАРК, р38 МАРК и Akt

3.2.6. Изучение влияния изопреналина и фармакологических ингибиторов ERK1,2 МАРК на буметанид-чувствительный транспорт калия и фосфорилирование ERK1,2 МАРК, р38 МАРК и

Akt в скелетных мышцах после однократного сеанса сократительной активности

3.4. Изучение фармакологической регуляции активности Na+-K+-2C1-котранспортера в "медленной" и "быстрой" скелетных мышцах крысы при инкубации в гиперосмолярной среде, полученной добавлением глюкозы или маннитола

3.4.1. Изучение влияния инсулина, изопреналина, форсколина и фармакологических ингибиторов ERK1,2 МАРК и р38 МАРК на буметанид-чувствительный транспорт Rb в скелетной мышце при инкубации в гиперосмолярной среде

3.4.2. Изучение влияния инсулина, изопреналина и форсколина на объём скелетной мышцы крысы при инкубации в гиперосмолярной среде

3.4.3. Изучение влияния инсулина и фармакологических ингибиторов ERK1,2 МАРК и р38 МАРК на фосфорилирование ERK1,2 МАРК, р38 МАРК и Akt при при инкубации скелетной мышцы в гиперосмолярной среде

3.5. Изучение роли протеин киназы А и АТФ-зависимых калиевых каналов в регуляции активности Ыа+-К+-2СГ-котранспортера в "медленной" и "быстрой" скелетных мышцах крысы 114 3.5.1. Изучение влияния фармакологических ингибиторов протеин киназы А и ERK1,2 МАРК на буметани д-чувствителы 1ый транспорт 86Rb и фосфорилирования ERK1,2 МАРК, р38 МАРК и

Akt в скелетной мышце

3.5.2. Изучение влияния гликолиза и АТФ-зависимых калиевых каналов в регуляции работы Ка+-К+-2СГ-котранспортера протеин киназой А и гиперосмолярной средой в скелетных мышцах крысы и

НЕК-293 клетках, имеющих высокую активность АТФ-зависимых калиевых каналов

3.6. Изучение действия фармакологических ингибиторов ERK1,2 МАРК, протеин киназ А и С и фосфатидилинозитол 3-киназы на активность Na+-K+-2Cr-KOTpaHcnopTepa в изолированной культуре скелетно-мышечных и кишечных эпителиальных клеток крысы

3.7. Изучение влияния длительного курса физических упражнений и сахарного диабета на активность Ка+-К+-2СГ-котранспортера в скелетной мышце крысы и человека

3.7.1. Изучение буметанид-чувствительного транспорта калия в "медленной" и "быстрой" скелетных мышц у здоровых крыс после двух- и четырёх-недельного бега на тредмиле. Влияние инсулина и изопреналина

3.7.2. Изучение экспрессии Ка+-К+-2СГ-котранспортера и фосфорилирования ERK1,2 МАРК и Akt в "медленной" и "быстрой" мышцах у здоровых крыс после двух- и четырёх-недельного бега на тредмиле. Влияние инсулина и изопреналина

3.7.3. Изучение экспрессии Ка+-К+-2СГ-котранспортера и фосфорилирования ERK1,2 МАРК и Akt в "медленной" и быстрой" мышцах у больных диабетом fa/fa Zucker крыс до и после лечения тиазолидинедионом (TZD) сахарного диабета. Влияние инсулина

3.7.4. Изучение экспрессии Ка+-К+-2СГ-котранспортера и фосфорилирования ERK1,2 МАРК и Akt в скелетной мышце у пациентов с сахарным диабетом до и после лечения сахарного диабета. Влияние инсулина

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391. Также выражаю искреннюю благодарность доктору Дональду Томасону за предоставленную помощь при проведении исследований.